Stem Cell Culture Media - Clinical grade

StemFit Basic03

Article number: Basic03

 

Benefits:

  • No animal- and human-derived components
  • Manufactured under strictly controlled conditions (GMP-compliant according to Japanese guidelines)
  • Media for embryonic stem cells and iPS cells
  • Save more than 50% of the media during cell cultivation
  • Reproducible and consistent growth rates
  • Higher proliferation rate
  • Available in Europe and in the US

 

Choose your media: Basic02 and Basic04 for basic research and Basic03 for clinical research

     

 

Stem cell medium for clinical research

StemFit® is a cell culture medium for clinical research into regenerative medicine,. This stem cell media is produced under strictly controlled conditions and is GMP compliant under Japansese guidlines. StemFit® Basic03 is completely free of animal and human components. All ingredients are recombinant.

 

 

Recommended by the nobel prize winner Shin’ya Yamanaka

“StemFit®, a newly developed Xf-medium, was the best medium for hESC and hiPSC culture with rLN511E8.” Download here the paper

 

Feeder cells and Xeno-free culture medium

StemFit® medium was developed to produce a reliable and well-defined growth condition for stem cells. It contains only Xeno-free components and all necessary components for the culture of embryonic as well as induced Pluripotent Stem (iPS) cells. These important benefits lead to a reduction of variances and contaminations in the cultivation of stem cells, when using the StemFit® medium.

 

Very reproducible growth rate

The cultivation of stem cells using StemFit® results in a very reproducible growth rate (Fig. 1). This enables a perfect planning of experiments. No more variation due to different starting conditions caused by the natural variation when culturing on feeder cells. Analysing the morphology of stem cells cultivated in StemFit® shows that the colony shape and size are very similar to the cells grown on feeder cells (Fig. 2).

Fig. 1: Reproducible and continuous growth rates of a human iPS-cell line. 201B7 hiPS cells were seeded at different concentrations. The cell number was determined every 7 days for four weeks.

Fig. 2: Colony morphology shows a very similar shape and size of the hiPS-cells when compared to feeder cells.

 

Less media volume needed

Due to the high-quality components and the ideal concentration of nutrients, the required volume of StemFit® is lower than of conventional media. Additionally, reduced feeding steps during the week, creates a total volume difference between StemFit® and conventional media of more than 50% (Fig 3.).

Fig. 3: The volume of StemFit® can be reduced to by 25% per well. The reduced amount of media changes leads to a further volume reduction of more than 50%.

 

Weekend-free feeding

The maintenance of stem cells is very complicated and labor intense and the feeding step during the weekends is very tedious. StemFit® allows a feeding-free weekend, by using the recommended weekly workflow shown below (Fig. 4).

 

Fig. 4: StemFit® allows a feeding-free weekend.

 

Start from a single cell

StemFit® enables a superior colony forming efficiency from a single cell clone, which minimize the effects of stress and result in reliable cells for downstream applications. Furthermore, with StemFit® you can easily determine the efficiency of your cell production and duplicate individual clones.

 

Easy transition from feeder cells

Majority of the stem cells are still maintained in feeder-cell dependent cultivation. The transition from feeder-cells to nonfeeder-cells system could not be easier: Switch the medium twice before passaging the cells and continue cultivating them in StemFit®. After 2-3 passages the feeder cells will be completely diluted out and a pure stem cell culture will be obtained (Fig. 5).

 

Fig. 5: Easy transition from feeder-cells to nonfeeder-cells systems with StemFit®.

 

Compatible with many matrices

StemFit® has been tested on many different matrices. As shown on Fig. 6., the fold expansion rate of cells cultured in StemFit® is much higher when compared to medium A from the market leading competitor.

Fig. 6: Different cell matrices were used as growing matrix and combined with StemFit® or with medium A. The fold expansion of 201B7 hiPS cells was determined after 7 days.

 

Consistent gene expression profile

The cultivation of stem-cells is very stressful for the cells. Every passage and growth period could therefore introduce unwanted changes in the genome expression profile. Hence, the CGI Catapult Institute in London investigated the genomic profile of StemFit® after 1 passage, 3 and 5 passages and compared it to 4 commercially available media. The most consistent gene expression was obtained using StemFit® (Fig. 7 , StemFit® labeled as AK03N).

Fig. 7: Consistent gene expression when using StemFit® Media (code name AK03N). The expression profile using the TaqMan ScoreCard™ assay (n=3) showed that the most consistent gene expression of after 5 passages were obtained using StemFit® media.

 

Less production of lactate

The production of lactate is the result of hypoxic stress. The consequences can be changes in the genome expression profile or lead to unwanted differentiation of the stem cells. The CGI Catapult Institute in London showed that there is considerably less lactate production when the cells are grown in StemFit® (Fig. 8).

Fig. 8: StemFit® Media (code name AK03N) caused a considerably lower accumulation of lactate when compared to 4 other commercially available Stem cell media.

 

 

 

References

 

StemFit® is recommended by leading Scientists

StemFit® medium was developed 2014 in Japan by the Aminoscience division at Ajinomoto. Since that time, StemFit® has been citied in more than 75 scientific publications so far! The responses from the scientific society, when usingusing the StemFit® media for embryonic stem cells and iPS cells, were very positive . Enclosed a citation from Nakagawa et al., 2014 (Yamanaka’s lab):

 

„A novel efficient feeder-free culture system for the derivation of human induced pluripotent stem cell„

 

StemFit®, a newly developed Xf-medium, was the best medium for hESC and hiPSC culture with rLN511E8.”

 

 

StemFit® is recommended by CiRA

The Centre for iPS Cell Research and Application has a central aim: Provide a better understanding of iPS cells and regenerative medicine, and to offer protocols for mouse and human derived iPS-cells. For their human iPS-cells protocol, they recommend to use StemFit®. They even wrote a paper showing the superior StemFit® features (Nakagawa et al., 2014). Here is a simplified procedure which is available on their website:

 

Citations:

Nakagawa, M. et al. (Yamanakas lab) (2014). A novel efficient feeder-free culture system for the derivation of human induced pluripotent stem cells. Scientific Reports, 4:3594.
Morizane R and Bonventre JV (2017). Generation of nephron progenitor cells and kidney organoids from human pluripotent stem cells. Nature Protocols, 12: 195-207.

Camp JG el al. (2017). Multilineage communication regulates human liver bud development from pluripotency. Nature, 456: 533-538.

Takamichi Miyazaki et al. (2017). Efficient Adhesion Culture of Human Pluripotent Stem Cells Using Laminin Fragments in an Uncoated Manner. Scientific Reports, 7: 41165.

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and many more…

 

Comparison Table for StemFit®

Basic02, Basic03 and Basic04

 

Stem cells successfully used with StemFit®:

(based from over 75 scientific reports)

 

Human embryonic stem cells (hESCs)Human induced pluripotent stem cells (hiPSCs)
253G11020C7
KhES1
1027B3
SEES-3
1027B6
SEES-4
1039A1
SEES-5
1120C12
SEES-6
1123C1-G
SEES-7
1156D15
1156D2
more comming soon...1210B2
1231A3
1335A1
1383D2
1383D6
1383D7
201B7
253G1
253G4
32R1
404C2
409B2
414C2
585A1
585B1
836B1
836B3
987A3
987A7
Ff-I06
Ff-I14
Ff-I01
MIFF3
TIG107
TkDA3-4
more comming soon...

Manual

StemFit Basic03_Manual

 

Technical Tips – Single-Cell cloning

StemFit Basic03_Single-Cell cloning

 

Technical Tips – High fold expansion

StemFit Basic03_High fold expansion

 

Can I use Stemfit in a bioreactor?

It is possible to use Basic02 in Bioreactors. There is a presentation deposit available.


How is the performance of StemFit in small clump splitting? Is it possile that the cell walls seem darker and the cell number is reduced?

We recommend single -cell passaging, but small clump passaging is also working. Cell morphologies will always vary among different media. Sometimes the edge of colonies seems “darker” if you use StemFit, but the morphology is always good. Regarding the cell number there are different possibilities:

1.: StemFit tends to make more packed colonies compared to other media, so the actual number would be larger than it’s surface confluency.

2.: In general, adaption to other medium requires several passages. Cell growth would be improved after 2-3 passages.


Which amount of StemFit should I use in a 12-well plate and which cell density is suitable?

In general the medium volume depends on the area of the culture vessels. Therefore, the volume of StemFit would be 750 µL and a cell density of 5,000 – 10,000 cells/well (Depending on the cell line).

However, the evaportaion is sometimes different between different sizes of plates. If cells grow poorly, we would recommend to increase the volume of media up to 1 ml.

3.: Addition of Rock inhibitor on the passaging day will increase cell viability and cell number.


Can I use StemFit for murine cells?

This is not possible, because murine cells require different types of growth factors compared to ES/iPSCs.

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